Studi perbandingan metode transformasi DNA menggunakan vektor Agrobacterium tumefaciens pada tanaman tebu (Sacharum hybrid)
In order to compare transient expression of gus gene driven by CaMV 35S and rice ubiquitin RUBQ2 promoters, a DNA transformation was conducted using embryogenic callus and suspension cultures of sugarcane. The transient gus expression was observed by histochemical staining method. The histochemica...
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| Format: | Article |
| Language: | English |
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Indonesian Biological Society
2012-02-01
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| Series: | Journal of Biological Researches |
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| Online Access: | http://berkalahayati.org/journal/5f8590cd |
| _version_ | 1818358766299512832 |
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| author | Sri Setyati Purnama Oktaviandari Muhammad Hazmi Bambang Sugiharto |
| author_facet | Sri Setyati Purnama Oktaviandari Muhammad Hazmi Bambang Sugiharto |
| author_sort | Sri Setyati |
| collection | DOAJ |
| description | In order to compare transient expression of gus gene driven by CaMV 35S and rice ubiquitin RUBQ2 promoters, a DNA
transformation was conducted using embryogenic callus and suspension cultures of sugarcane. The transient gus expression was
observed by histochemical staining method. The histochemical observation of GUS activity after co-cultivation showed that RUBQ2
promoter produced high level of clear blue spots both in embryogenic callus and suspension cultures, while the CaMV35S promoter
was not detected. The suspension cultures slightly increased transient gus gene expression compared to embryogenic callus. However,
the histochemical analysis of regenerated putative transformant plants after 5 successive cycles on the selection medium showed no
blue spots of gus gene expression. PCR amplification of DNA for CaMV35 or nptII in putative transformant plants confirmed that there
was no integration of the transformed gene in the genome DNA. The results suggested a possibility of somaclonal variation with callus
propagation, thus did not produce transformed plants. To avoid the somaclonal variation, the transformation was conducted using in
vitro plants and multiple shoots without intervening callus phase. Histochemical observation of infected materials after co-cultivation
showed that almost all of the infected materials partially exhibited blue color in the basal region. In case of in vitro plants, they rapidly
grow and multiplied in the selection medium, thus the method provided an excellent system for the transformation in sugarcane. The
results suggest that in vitro plants as well as multiple shoots need further investigation to be used as target tissues for Agrobacteriummediated
transformation in sugarcane. |
| first_indexed | 2024-12-13T20:34:13Z |
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| id | doaj.art-87c211c2e1b845bc86c3fa56704f75e2 |
| institution | Directory Open Access Journal |
| issn | 0852-6834 2337-389X |
| language | English |
| last_indexed | 2024-12-13T20:34:13Z |
| publishDate | 2012-02-01 |
| publisher | Indonesian Biological Society |
| record_format | Article |
| series | Journal of Biological Researches |
| spelling | doaj.art-87c211c2e1b845bc86c3fa56704f75e22022-12-21T23:32:20ZengIndonesian Biological SocietyJournal of Biological Researches0852-68342337-389X2012-02-01131394410.23869/bphjbr.13.1.20076Studi perbandingan metode transformasi DNA menggunakan vektor Agrobacterium tumefaciens pada tanaman tebu (Sacharum hybrid)Sri Setyati0Purnama Oktaviandari1Muhammad Hazmi2Bambang Sugiharto3Pusat Penelitian Biologi Molekul dan *** Fakultas Matematika dan Pengetahuan Alam Universitas Jember ** Fakultas Pertanian Universitas Muhammadiyah JemberPusat Penelitian Biologi Molekul dan *** Fakultas Matematika dan Pengetahuan Alam Universitas Jember ** Fakultas Pertanian Universitas Muhammadiyah JemberPusat Penelitian Biologi Molekul dan *** Fakultas Matematika dan Pengetahuan Alam Universitas Jember ** Fakultas Pertanian Universitas Muhammadiyah JemberPusat Penelitian Biologi Molekul dan *** Fakultas Matematika dan Pengetahuan Alam Universitas Jember ** Fakultas Pertanian Universitas Muhammadiyah JemberIn order to compare transient expression of gus gene driven by CaMV 35S and rice ubiquitin RUBQ2 promoters, a DNA transformation was conducted using embryogenic callus and suspension cultures of sugarcane. The transient gus expression was observed by histochemical staining method. The histochemical observation of GUS activity after co-cultivation showed that RUBQ2 promoter produced high level of clear blue spots both in embryogenic callus and suspension cultures, while the CaMV35S promoter was not detected. The suspension cultures slightly increased transient gus gene expression compared to embryogenic callus. However, the histochemical analysis of regenerated putative transformant plants after 5 successive cycles on the selection medium showed no blue spots of gus gene expression. PCR amplification of DNA for CaMV35 or nptII in putative transformant plants confirmed that there was no integration of the transformed gene in the genome DNA. The results suggested a possibility of somaclonal variation with callus propagation, thus did not produce transformed plants. To avoid the somaclonal variation, the transformation was conducted using in vitro plants and multiple shoots without intervening callus phase. Histochemical observation of infected materials after co-cultivation showed that almost all of the infected materials partially exhibited blue color in the basal region. In case of in vitro plants, they rapidly grow and multiplied in the selection medium, thus the method provided an excellent system for the transformation in sugarcane. The results suggest that in vitro plants as well as multiple shoots need further investigation to be used as target tissues for Agrobacteriummediated transformation in sugarcane.http://berkalahayati.org/journal/5f8590cdDNA transformationAgrobacterium tumefaciensrice ubiquitin promoterCaMV35S promotersugarcane |
| spellingShingle | Sri Setyati Purnama Oktaviandari Muhammad Hazmi Bambang Sugiharto Studi perbandingan metode transformasi DNA menggunakan vektor Agrobacterium tumefaciens pada tanaman tebu (Sacharum hybrid) Journal of Biological Researches DNA transformation Agrobacterium tumefaciens rice ubiquitin promoter CaMV35S promoter sugarcane |
| title | Studi perbandingan metode transformasi DNA menggunakan vektor Agrobacterium tumefaciens pada tanaman tebu (Sacharum hybrid) |
| title_full | Studi perbandingan metode transformasi DNA menggunakan vektor Agrobacterium tumefaciens pada tanaman tebu (Sacharum hybrid) |
| title_fullStr | Studi perbandingan metode transformasi DNA menggunakan vektor Agrobacterium tumefaciens pada tanaman tebu (Sacharum hybrid) |
| title_full_unstemmed | Studi perbandingan metode transformasi DNA menggunakan vektor Agrobacterium tumefaciens pada tanaman tebu (Sacharum hybrid) |
| title_short | Studi perbandingan metode transformasi DNA menggunakan vektor Agrobacterium tumefaciens pada tanaman tebu (Sacharum hybrid) |
| title_sort | studi perbandingan metode transformasi dna menggunakan vektor agrobacterium tumefaciens pada tanaman tebu sacharum hybrid |
| topic | DNA transformation Agrobacterium tumefaciens rice ubiquitin promoter CaMV35S promoter sugarcane |
| url | http://berkalahayati.org/journal/5f8590cd |
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